Molecular fluorescence spectroscopy

The basic design of instrumentation for monitoring molecular fluorescence and molecular phosphorescence is similar to that found for other spectroscopies. The most significant differences are discussed in the following sections. 

Molecular Fluorescence A typical instrumental block diagram for molecular fluorescence is shown in Figure 10.45. In contrast to instruments for absorption spectroscopy, the optical paths for the source and detector are usually positioned at an angle of 90°. 

Molecular Fluorescence Spectroscopy Photometric Titrations Analytical Applications of Interferometry Vol. 9 Ultraviolet Photoelectron and Photoion Spectroscopy... 

McGown, L.B., and Warner, I.M. (1990) Molecular fluorescence, phosphorescence, and chemiluminescence spectroscopy. Anal. Chem. 190, 255R. 

Berberan-Santos M. N. (2001) Pioneering Contributions of Jean and Francis Perrin to Molecular Fluorescence, in Valeur B. and Brochon J. C. (Eds), New Trends in Fluorescence Spectroscopy. 

Although this book is devoted to molecular fluorescence in condensed phases, it is worth mentioning the relevance of fluorescence spectroscopy in supersonic jets (Ito et al., 1988). A gas expanded through an orifice from a high-pressure region into a vacuum is cooled by the well-known Joule-Thomson effect. During expansion, collisions between the gas molecules lead to a dramatic decrease in their translational velocities. Translational temperatures of 1 K or less can be attained in this way. The supersonic jet technique is an alternative low-temperature approach to the solid-phase methods described in Section 3.5.2 all of them have a common aim of improving the spectral resolution. 

Tunnelling electrons from a STM have also been used to excite photon emission from individual molecules, as has been demonstrated for Zn(II)-etioporphyrin I, adsorbed on an ultrathin alumina film (about 0.5 nm thick) grown on a NiAl(l 10) surface (Qiu et al, 2003). Such experiments have demonstrated the feasibility of fluorescence spectroscopy with submolecular precision, since hght emission is very sensitive to tip position inside the molecule. As mentioned before the oxide spacer serves to reduce the interaction between the molecule and the metal. The weakness of the molecule-substrate interaction is essential for the observation of STM-excited molecular fluorescence. 

Aznarez et al. [2] also applied molecular fluorescence spectroscopy to the determination of borate in soils. The boron is extracted from the soil with 2-methylpentane-2,4 diol into methyl isobutyl ketone and 0.1% m/v diben-zoylmethane in methyl isobutyl ketone and concentrated phosphoric acid is... 

The identification and quantitative determination of specific organic compounds in very complex samples is an area of intense current research activity in analytical chemistry Optical spectroscopy (particularly UV-visible and infrared absorption and molecular fluorescence and phosphorescence techniques) has been used widely in organic analysis. Any optical spectroscopic technique to be used for characterization of a very complex sample, such as a coal-derived material, should exhibit very high sensitivity (so that trace constituents can be determined) and extremely great selectivity (so that fractionation and separation steps prior to the actual analysis can be held to the minimum number and complexity). To achieve high analytical selectivity, an analytical spectroscopic technique should produce highly structured and specific spectra useful for "fingerprinting purposes," as well as to minimize the extent of overlap of spectral bands due to different constituents of complex samples. 

The technique of matrix isolation has been shown to produce highly characteristic spectra of individual components of complex samples combination of MI spectroscopy with separation techniques promises to increase further the analytical capabilities of the technique. While our research to date has emphasized FTIR and molecular fluorescence spectrometry, MI as a sampling procedure is not limited to these two forms of spectrometry. For example, some interesting preliminary analytical results by MI Raman spectrometry recently have been described (32). It should also be stressed that the cryogenic procedures required for the vast majority of MI spectral studies are neither difficult nor unduly expensive except in very special cases, closed-cycle cryostats requiring no cryogenic liquids (and no prior experience in low-temperature techniques) are entirely satisfactory for MI... 

Molecular fluorescence spectroscopy is a commonly employed analytical method that is sensitive to certain chemical properties of FA (9-13). Fulvic acid s molecular fluorescence is principally due to conjugated unsaturated segments and aromatic moieties present in the macromolecule (14). Several types of fluorescence spectra can be measured, including an excitation emission matrix or total luminescence spectrum, constant offset synchronous fluorescence, excitation spectra, and emission spectra, furnishing the researcher with useful data. The ability to resolve and select multiple fluorescent species makes these approaches extremely useful for studying FA relative to its chemical reactivity. 

Size exclusion chromatography (SEC) has been used to measure molecular weight (MW) distribution of humic substances (3, 6-9). Coupled with detection methods such as molecular fluorescence spectroscopy and dissolved organic carbon analysis (7), electrochemical detection (9), and atomic emission spectroscopy (5), SEC has been used extensively to study humic-metal complexes. A major disadvantage of SEC is that it does not provide adequate resolution for separating humic materials as they do not appear to be made up of distinct fractions with large differences in MW. 

Information about conformation and motion of nucleic acids was obtained by using pulsed fluorescence and fluorescence correlation spectroscopy 25,47). A structural conformation can often be fingerprinted by the fluorescence lifetime of molecular fluorescing group or of an inserted label. 

Fluorescence. The use of molecular fluorescence spectroscopy for the quantitation of enzyme reaction products has resulted in detection limits that are several orders of magnitude lower than those achieved by standard absorbance methods. At low analyte concentrations, fluorescence emission intensity is directly proportional to concentration, and its value depends on both the molar absorptivity of the analyte at the excitation wavelength, and the fluorescence quantum yield of the analyte, under the assay conditions. 

In this chapter, we discuss the basic principles that are necessary to understand measurements made with electromagnetic radiation, particularly those deeding with the absorption of UV, visible, and IR radiation. The nature of electromagnetic radiation and its interactions with matter are stressed. The next four chapters are devoted to. spectroscopic instruments (Chapter 25), molecular absorption spectroscopy (Chapter 26), molecular fluorescence spectroscopy (Chapter 27), and atomic spectroscopy (Chapter 28). 

Lasing can be sustained over a continuous range of wavelengths on the order of 40 to 50 nm. The broad band over which lasing occurs makes the dye laser suitable for tuning by inserting a grating, a filter, a prism, or an interferometric element into the laser cavity. Dye lasers are very useful for molecular fluorescence spectroscopy and many other applications. 

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